LOCUS       sequence-160930-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p63 (580 a.a.) tagged with mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  AUTHORS   van Unen J, Yin T, Wu YI, Mastop M, Gadella TW Jr, Goedhart J
  TITLE     Kinetics of recruitment and allosteric activation of ARHGEF25 
            isoforms by the heterotrimeric G-protein Galphaq.
  JOURNAL   Sci Rep. 2016 Nov 11;6:36825. doi: 10.1038/srep36825.
  PUBMED    27833100
REFERENCE   2  (bases 1 to 6463)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-11-11"; volume: "6:36825. doi"; pages: " 
            10.1038/srep36825"
COMMENT     Sequence Label: p63RhoGEF580-mVenus-N1
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             654..740
                     /codon_start=1
                     /product="palmitoylation signal consisting of the first 29 
                     amino acids of p63RhoGEF (Goedhart et al., 2013)"
                     /label=palmitoylation signal
                     /translation="MRGGHKGGRCACPRVIRKVLAKCGCCFAR"
     regulatory      1215..1224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2403..2412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2409..2411,2412..2414,2415..3128)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2454..2475)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2715..2734)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3062..3083
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3251..3372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3288..3307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3342..3361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3379..3834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3516..3537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3728..3747
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3861..3965
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3967..4324
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3967..4158
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3990..4010)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4175..4310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4359..5153
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4359..5153
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4413..4432)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5023..5042
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5341..5360)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5385..5432
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5761..6349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6250..6269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc accatgcggg
      661 gggggcacaa agggggtcgc tgtgcctgtc cccgtgtgat ccgaaaagtg ctggcaaaat
      721 gcggctgctg cttcgcccgg gggggacgtg aatcctattc cattgcgggc agtgagggga
      781 gtatatcggc ttctgctgcc tccggtctgg ctgccccctc tggccccagc tctggcctca
      841 gctctggccc ctgttcccca ggccccccag ggcccgtcag tggcctgagg agatggttgg
      901 atcattccaa acattgtctc agtgtggaaa ctgaggcaga cagtggtcag gcaggaccat
      961 atgagaactg gatgttggag ccagctctag ccacaggaga ggagctgccg gaactgacct
     1021 tgctgaccac actgttggag ggccctggag ataagacgca gccacctgaa gaggagactt
     1081 tgtcccaagc ccctgagagt gaggaggaac agaagaagaa ggctctggaa aggagtatgt
     1141 atgtcctgag tgaactggta gaaacagaga aaatgtacgt ggacgacttg gggcagattg
     1201 tggagggtta tatggccacc atggctgctc agggggtccc cgagagtctt cgaggccgtg
     1261 acaggattgt gtttgggaat atccagcaaa tctatgagtg gcaccgagac tatttcttgc
     1321 aagagctaca acggtgtctg aaagatcctg attggctggc tcagctattc atcaaacacg
     1381 agcgccggct gcatatgtat gtggtgtact gtcagaataa gcccaagtca gagcatgtgg
     1441 tgtcagagtt tggggacagc tactttgagg agctccggca gcagctgggg caccgcctgc
     1501 agctgaacga cctcctcatc aaacctgtgc agcggatcat gaaataccag ctgctgctca
     1561 aggattttct caagtattac aatagagctg ggatggatac tgcagaccta gagcaagctg
     1621 tggaggtcat gtgctttgtg cccaagcgct gcaacgatat gatgacgctg gggagattgc
     1681 ggggatttga gggcaaactg actgctcagg ggaagctctt gggccaggac actttctggg
     1741 tcaccgagcc tgaggctgga gggctgctgt cttcccgagg tcgagagagg cgcgtcttcc
     1801 tctttgagca aatcatcatc ttcagtgaag ccctgggagg aggagtgaga ggtggaacac
     1861 agcctggata tgtatacaag aacagcatta aggtgagctg cctgggactg gaggggaacc
     1921 tccaaggtga cccttgccgc tttgcactga cctccagagg gccagagggt gggatccagc
     1981 gctatgtcct gcaggctgca gaccctgcta tcagtcaggc ctggatcaag catgtggctc
     2041 agatcttgga gagccaacgg gacttcctca acgcattgca gtcacccatt gagtaccaga
     2101 gacgggagag ccagaccaac agcctggggc ggccaagagg gcctggagtg gggagccctg
     2161 gaagaattcg gcttggagat caggcccagg gcagcacaca cacacccatc aatggctctc
     2221 tcccctctct gctgctgtca cccaaagggg aggtggccag agccctcttg ccactggata
     2281 aacaggccct tggtgacatc ccccaggctc cccatgactc tcctccagtc tctccaactc
     2341 caaaaacccc tccctgccaa gccagacttg ccaagctgga tgaagatgag ctgtcaccgg
     2401 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2461 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     2521 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     2581 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
     2641 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2701 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2761 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2821 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     2881 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     2941 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3001 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     3061 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3121 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3181 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3241 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3301 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3361 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3421 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3481 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3541 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3601 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3661 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3721 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3781 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3961 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4081 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4141 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4201 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4261 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4321 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4381 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4441 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4501 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4561 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4621 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4681 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4741 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4801 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4861 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4921 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4981 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5041 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5101 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5161 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5221 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5281 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5341 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5401 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5461 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5521 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5581 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5641 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5881 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6001 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6061 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6421 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
